The mechanical properties of electrospun fiber networks are critical in a range of applications from filtration to tissue engineering and are dependent on the adhesion between contacting fibers within the network. This adhesion is complex as electrospun networks exhibit a variety of contacts, including both cross-cylinder and parallel fiber configurations. In situ atomic force microscopy (AFM) was used to quantify the work of adhesion between a pair of individual electrospun polyamide fibers using controlled orientations and measurable contact areas. The work of adhesion was found to depend strongly on the fiber-fiber contact, with the separation of fibers in a parallel fiber configuration exhibiting considerably higher work of adhesion across a range of contact lengths than a cross-cylinder configuration. Our work therefore highlights direction-dependent adhesion behavior between electrospun fibers due to a suggested polymer chain orientation mechanism which increases net van der Waals interactions and indicates the variability of adhesion within a random electrospun fiber network.